Journal: Obesity Science & Practice

Article Title: A Cross‐Sectional Study Investigating the Relationship Between the FTO Gene Polymorphism in Relation to Obesity Traits and Vitamin D Status in Adolescence

doi: 10.1002/osp4.70112

Figure Lengend Snippet: Bar chart showing the percentage distribution of FTO genotypes (TT, TA, AA) within each combined BMI category (Underweight/Normal and Overweight/Obese).

Article Snippet: Genotyping the FTO rs9939609 was performed using TaqMan genotyping assays by allelic discrimination (assay ID: C__30090620_10) according to Life Technologies protocols (Thermo Fisher Scientific, MA, USA) as previously published [ ].

Techniques:

Journal: Cell Biology and Toxicology

Article Title: FTO-mediated m6A demethylation of CSF3 suppresses NETosis via downregulation of RLN2 expression in colorectal cancer

doi: 10.1007/s10565-025-10120-9

Figure Lengend Snippet: FTO mediates m6A demethylation of CSF3 and inhibits its expression. ( A ) The colorimetric m6A detection assay showed the overall m6A levels in CRC cells. ( B ) MeRIP combined with specific qRT-PCR was utilized to detect the relative m6A enrichment of CSF3 transcript in CCD 841 CoN and CRC cells. ( C ) Protein expression of FTO detected by western blot assay. ( D ) Transfection efficiency of oe-FTO measured in CRC cells. ( E ) qRT-PCR was utilized to detect the mRNA expression of CSF3 in CRC cells. ( F ) The colorimetric m6A detection assay showed the overall m6A levels in CRC cells after upregulation of FTO. ( G ) MeRIP combined with specific qRT-PCR was utilized to detect the relative m6A enrichment of CSF3 transcript in CCD 841 CoN and CRC cells after upregulation of FTO. ( H ) RIP assay using an anti-FTO antibody or IgG to detect the binding to CSF3 in CRC cells. IgG was used as the negative control. *compared with CCD 841 CoN or oe-NC or IgG groups. N = 3 wells per group

Article Snippet: Subsequently, sections were probed with primary antibodies including FTO (1:200, #27,226–1-AP, proteintech) and Ki-67 (1:1000, #28,074–1-AP, proteintech) at 4°C overnight.

Techniques: Expressing, Detection Assay, Quantitative RT-PCR, Western Blot, Transfection, Binding Assay, Negative Control

Journal: Cell Communication and Signaling : CCS

Article Title: Deregulation of m6A-RNA methylation impairs adaptive hypertrophic response and drives maladaptation via mTORC1-S6K1-hyperactivation and autophagy impairment

doi: 10.1186/s12964-025-02509-0

Figure Lengend Snippet: Cardiomyocyte specific FTO deficiency causes early dilatation and reduces compensatory hypertrophy. Two-month-old FTOcKO and Cre control animals were analyzed one week after TAC surgery using 26-G needle A - I . The Echocardiography is measured for each group: Cre control in blue (sham & TAC) and FTOcKO in red (sham and TAC); ( A ) The Ejection fraction (EF) and ( B ) Fractional Shortening (FS) which measures the cardiac function are presented in percentage. C LVID (left ventricular interdimensional diameter) is measured in mm; ( D ) Intraventricular septum thickness is measured in mm; ( E ) The LV posterior wall thickness (LVPW) is measured in mm; ( F ) The ratio of LV weight-to- tibia length (LV/TL in mg/mm) is calculated for each mice; ( G ) The relative wall thickness (RWT) is calculated by dividing the sum of anterior wall thickness (LVAW) and posterior wall thickness (LVPW) by the LV inner dimension (LVID) ((LVAW + LVPW)/LVID); ( H ) representative transverse sections of FTOcKO and Control mice stained using wheat germ agglutinin (WGA); ( I ) The cross-sectional area for the WGA stained sections are represented in µm 2 ; ( J ) representative transverse sections of FTOcKO and Control mice stained using picro sirius stain kit; ( K ) the fibrotic area is represented in percentage; Each dot represents the measurement of one animal, significance levels presented using ordinary two-way ANOVA with Tukey’s correction for multiple comparison analysis. The mean with SEM is presented; mice n for A-G (CreC Sham, n = 3–6; CreC TAC, n = 5–11; FTOcKO Sham, n = 3–4; FTOcKO TAC, n = 7–8); mice n for I (CreC Sham, n = 3; CreC TAC, n = 3; FTOcKO Sham, n = 4; FTOcKO TAC, n = 3); mice n for K (CreC sham, n = 3; CreC TAC, n = 4; FTOcKO Sham, n = 3; FTOcKO TAC, n = 3). For hypertrophic analysis in vitro, hiPS-CMs were transfected with scr siRNAs (scr) or with FTO siRNAs (siFTO) and their hypertrophic response were studied by treating the silenced CMs with 3 nM of ET-1 (Endothelin-1) for 24 h; ( L ) show the representative images of scr and siFTO transfected CMs with or without ET-1. Green fluorescence represents the alpha-actinin (sarcomeric) and blue is stained with DAPI; ( M ) represents the hypertrophic measurement and the relative cell size for each CM is measured using ImageJ. The mean with SEM is presented. n represents cells per dish (Scr, n = 5; Scr + ET-1, n = 4; siFTO, n = 5; siFTO + ET-1, n = 5); ordinary two way-ANOVA with Tukey`s correction for multiple comparison was performed and significance levels were shown for each condition in comparison to the control. *** P < 0.001; **** p < 0.0001; ** p < 0.005; * p < 0.05

Article Snippet: The Taqman Universal PCR master mix (Cat. No. 4304437) and the specific qPCR primers for AKT1S1 (TaqMan Gene expression Assay ID Hs00982883_m1; AKT1S1); TFEB ( TaqManTM Gene Expression Assay Hs00292981_m1; TFEB); and FTO (Taqman Gene expression Assay ID Hs01057143_m1; FTO) were purchased from Invitrogen.

Techniques: Control, Staining, Comparison, In Vitro, Transfection, Fluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Deregulation of m6A-RNA methylation impairs adaptive hypertrophic response and drives maladaptation via mTORC1-S6K1-hyperactivation and autophagy impairment

doi: 10.1186/s12964-025-02509-0

Figure Lengend Snippet: Differentially m6A methylated transcripts of FTO ablated animals. The MeRIP was performed for the heart tissues of FTOcKO and Cre control groups, hearts were isolated one week-post TAC surgery; ( A ) Detailed representation of differentially methylated transcript for each condition, CreC (sham& TAC) and FTOcKO (Sham &TAC), with FC1.5 FDR 0.05. (CreC: Cre Control group); ( B ) Gene Ontology enrichment of differentially methylated transcripts, FTOcKO Vs Cre control, with FC > 1.2 FDR 0.05: the biological process of differentially hypomethylated transcripts and differentially hypermethylated transcripts; ( C ) Biological pathways corresponding to differentially methylated transcripts in FTOcKO TAC group compared to CreC TAC mice groups

Article Snippet: The Taqman Universal PCR master mix (Cat. No. 4304437) and the specific qPCR primers for AKT1S1 (TaqMan Gene expression Assay ID Hs00982883_m1; AKT1S1); TFEB ( TaqManTM Gene Expression Assay Hs00292981_m1; TFEB); and FTO (Taqman Gene expression Assay ID Hs01057143_m1; FTO) were purchased from Invitrogen.

Techniques: Methylation, Control, Isolation

Journal: Cell Communication and Signaling : CCS

Article Title: Deregulation of m6A-RNA methylation impairs adaptive hypertrophic response and drives maladaptation via mTORC1-S6K1-hyperactivation and autophagy impairment

doi: 10.1186/s12964-025-02509-0

Figure Lengend Snippet: FTO ablation in the heart triggers hypermethylation in key transcripts of apoptosis inhibition, mTORC1-S6K1 signaling and autophagy: The transcription level m6A methylation changes are shown under conditions FTOcKO TAC vs CreC TAC for the transcripts ( A ) CFLAR, ( B ) CDK1, ( C ) AKT1S1 and ( D ) TFEB; MeRIP-qPCR was performed for the transcripts AKT1S1 and TFEB in scr and FTO-silenced hiPS-CMs, ( E ) shows the m6A enrichment of AKT1S1 and ( F ) shows the m6A enrichment of TFEB in MeRIP-siFTO compared to MeRIP-Scr, IgG Scr and IgG siFTO are the controls for m6A RNA immunoprecipitation; ( G ) mRNA expression data of FTO gene in Scr and siFTO hiPS-CMs; ( H ) relative gene expression of AKT1S1 in scr and siFTO hiPS-CMs; ( I ) relative gene expression of TFEB in scr and siFTO hiPS-CMs; ( J ) representative western blot image showing the signals of PRAS40 and TFEB proteins in Scr and siFTO-hiPS-CMs; ( K ) and ( L ) shows the pooled quantitative densitometry analysis for PRAS40 and TFEB respectively. The mean with SEM is presented for G-I and K-L, n represents no. of biological replicates; n = 4–6 for scr and n = 4–6 for siFTO hiPS-CMs; * p < 0.05; ** p < 0.005; *** < 0.001 all by two tailed unpaired t-test with Welch’s correction

Article Snippet: The Taqman Universal PCR master mix (Cat. No. 4304437) and the specific qPCR primers for AKT1S1 (TaqMan Gene expression Assay ID Hs00982883_m1; AKT1S1); TFEB ( TaqManTM Gene Expression Assay Hs00292981_m1; TFEB); and FTO (Taqman Gene expression Assay ID Hs01057143_m1; FTO) were purchased from Invitrogen.

Techniques: Inhibition, Methylation, RNA Immunoprecipitation, Expressing, Gene Expression, Western Blot, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Deregulation of m6A-RNA methylation impairs adaptive hypertrophic response and drives maladaptation via mTORC1-S6K1-hyperactivation and autophagy impairment

doi: 10.1186/s12964-025-02509-0

Figure Lengend Snippet: Loss of FTO demethylase induces cardiac apoptosis: ( A ) Representative western blots of Cre Control and FTOcKO heart samples with sham and TAC surgery. The apoptotic markers cleaved caspase 3 (cl.Casp3) and cleaved PARP (cl.PARP) were increased in FTOcKO Sham and TAC; ( B ) & ( C ) are pooled quantitative densitometry analysis for apoptotic markers cl.casp3 and cl.PARP respectively, from the heart samples of FTOcKO and Cre control groups (Sham &TAC); The mean with SEM is presented; n is no. of mice (CreC Sham, n = 4–5; CreC TAC, n = 5; FTOcKO Sham, n = 5; FTOcKO TAC, n = 5); ( D ) representative images of TUNEL staining of left ventricle (LV) sections from FTOcKO and Cre control animals (both Sham and TAC); ( E ) pooled TUNEL analysis, the percentage of TUNEL positive cells from the total number of cardiomyocytes were calculated; The mean with SEM is presented, mice condition (CreC Sham, n = 3; CreC TAC, n = 3; FTOcKO Sham, n = 3; FTOcKO TAC, n = 3); Statistical analysis ( B , C , E ) is by ordinary two-way ANOVA with Tukey’s correction for multiple comparisons; ( F ) Representative western blots of hiPS-CMs transfected with scr and FTO siRNAs; and the pooled densitometry analysis for cl.casp3 ( G ) and cl.PARP ( H ) were shown. I fluorescent double staining of alpha-actinin and TUNEL in hiPS-CMs with scr and siFTO silencing, and ( J ) represents the percentage of TUNEL positive cells (pooled data). The mean with SEM is presented. For G, H, J, biological replicate, n = 3–6 for each condition; For G & H, 1-sample t-test is performed, the results were normalized to the respective control conditions and ratios, in which a normal distribution of results cannot be proven, were analyzed. * p < 0.05; ** p < 0.005 all by unpaired t-test

Article Snippet: The Taqman Universal PCR master mix (Cat. No. 4304437) and the specific qPCR primers for AKT1S1 (TaqMan Gene expression Assay ID Hs00982883_m1; AKT1S1); TFEB ( TaqManTM Gene Expression Assay Hs00292981_m1; TFEB); and FTO (Taqman Gene expression Assay ID Hs01057143_m1; FTO) were purchased from Invitrogen.

Techniques: Western Blot, Control, TUNEL Assay, Staining, Transfection, Double Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Deregulation of m6A-RNA methylation impairs adaptive hypertrophic response and drives maladaptation via mTORC1-S6K1-hyperactivation and autophagy impairment

doi: 10.1186/s12964-025-02509-0

Figure Lengend Snippet: FTO depletion in cardiomyocytes significantly increased the mTORC1 signaling pathway: Representative western blots of Cre Control and FTOckO animals with Sham and TAC surgery. A The phosphorylation of S6 (ser235/236), ( B ) p70S6 kinase (thr389) and ( C ) 4EBP1 (Thr37/46) were increased in FTOcKO mice groups; pooled quantitative densitometry analysis of P-S6 ( D ), P-S6K1 ( E ), P-4EBP1 ( F ); The mean with SEM is presented; mice n = 4 for each condition; Statistical analysis ( D , E , F ) is by ordinary two-way ANOVA with Tukey’s correction for multiple comparisons; * p < 0.05, ** p < 0.005; ( G ) representative P-S6 (green) staining of FTOcKO and Cre control LV sections; ( H ) the percentage of p-S6 signal per field is analysed in CreC and FTOckO LV sections; The mean with SEM is presented; n represents each LV sections, n = 7–8 from 3 different mice; statistical analysis is by unpaired t-test with Welch’s correction, **** p < 0.0001; ( I ) Representative western blots of hiPS-CMs transfected with scr and siFTO; ( J ), ( K ) & ( L ) shows the pooled quantitative densitometry analysis of P-S6K1, P-S6 and P-4EBP1 respectively; ( M ) Fluorescent double staining of P-S6 (red) and alpha-actinin(sarcomeric) (green). The mean with SEM is presented; biological replicate, n = 3 for each condition; the results were normalized to the respective control conditions and ratios, in which a normal distribution of results cannot be proven, were analyzed * p < 0.05 all by unpaired t-test with Welch’s correction for J-L

Article Snippet: The Taqman Universal PCR master mix (Cat. No. 4304437) and the specific qPCR primers for AKT1S1 (TaqMan Gene expression Assay ID Hs00982883_m1; AKT1S1); TFEB ( TaqManTM Gene Expression Assay Hs00292981_m1; TFEB); and FTO (Taqman Gene expression Assay ID Hs01057143_m1; FTO) were purchased from Invitrogen.

Techniques: Western Blot, Control, Phospho-proteomics, Staining, Transfection, Double Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Deregulation of m6A-RNA methylation impairs adaptive hypertrophic response and drives maladaptation via mTORC1-S6K1-hyperactivation and autophagy impairment

doi: 10.1186/s12964-025-02509-0

Figure Lengend Snippet: FTO depletion and inhibition of late autophagy exacerbates cardiac apoptosis and impairs autophagic flux: The hiPS-CMs were transfected with siRNAs of FTO and scr, and the late-autophagy (fusion of autophagosomes and lysosomes) were blocked by using the inhibitors, Bafilomycin A (Baf) and Chloroquine (CQ) with the final concentration of 50 nM and 50 uM, respectively for 16 h; ( A ) Representative western blots of the autophagic markers (LC3 II and P62) and apoptosis (cl.PARP); pooled quantitative densitometry analysis of cl.PARP ( B ), P62 ( C ) and LC3 II ( D ); ( E ) Representative fluorescent images of LC3 II and p62 co-localization (P62-red; LC3 II – green; DAPI-blue) for scr and siFTO iPS-CMs treated with or without 50uM of Chloroquine (CQ) for 16 h. The mean with SEM is presented. biological replicate, n = 3–5 for each condition; the results were normalized to the respective control conditions and ratios, in which a normal distribution of results cannot be proven, were analyzed ** p < 0.005; * p < 0.05 all by unpaired t-test; ( F ) represents the live imaging of tandem mRFP-GFP-LC3 assay, the hiPS-CMs were transduced with RP-LC3 for 24 h and further transfected with scr and FTO siRNAs before treatment with or without Baf for 12 h; ( G ) represents the quantified data of LC3 puncta per cell under each conditions (Scr, siFTO, Scr + Baf and siFTO + Baf), black indicates total LC3 puncta per cell; yellow represents no. of autophagosomes per cell and red represents no. of autolysosomes per cell; total of 15 to 30 cells were analysed per dish, in total 6 dishes were considered for each condition. The mean value of all the cells for each condition were plotted in graph

Article Snippet: The Taqman Universal PCR master mix (Cat. No. 4304437) and the specific qPCR primers for AKT1S1 (TaqMan Gene expression Assay ID Hs00982883_m1; AKT1S1); TFEB ( TaqManTM Gene Expression Assay Hs00292981_m1; TFEB); and FTO (Taqman Gene expression Assay ID Hs01057143_m1; FTO) were purchased from Invitrogen.

Techniques: Inhibition, Transfection, Concentration Assay, Western Blot, Control, Imaging, Transduction

Journal: Cell Communication and Signaling : CCS

Article Title: Deregulation of m6A-RNA methylation impairs adaptive hypertrophic response and drives maladaptation via mTORC1-S6K1-hyperactivation and autophagy impairment

doi: 10.1186/s12964-025-02509-0

Figure Lengend Snippet: FTO ablation induces apoptosis and atrophy via mTORC1-S6K1 pathway: The hiPS-CMs were transfected with siRNAs of FTO and scr for 24 h, further treated with mTORC1 inhibitors Rapamycin (Rapa) and PF-4709671(S6Ki) with 50 nM and 5 µM final concentration respectively, for another 24 h: ( A ) Representative images of TUNEL staining for each condition (Un – Untreated); ( B ) pooled TUNEL analysis, the percentage of TUNEL positive cells from the total number of cardiomyocytes were calculated for each condition, biological replicate, n = 3–4 for each condition; ( C ) Representative western blots of FTO silenced hiPS-CMs with or without mTORC1 inhibitors, the activity of mTORC1-S6K1 is showed by P-S6 signals; cl.PARP signal represents cardiac apoptosis; ( D ) pooled quantitative densitometry analysis of cl.PARP (marker of apoptosis); biological replicate, n = 5 for each condition; * p < 0.05 and ** p < 0.01 all by student’s t-test; ( E - I ) Study of the effect of hypertrophic response by Endothelin-1(ET-1) in siFTO-hiPS-CMs and scr hiPS-CMs with or without PF-4709671 (S6K1i); 24 h after transfection, the CMs were incubated with or without 10 nM of ET-1 and 5 µM of S6K1i for 24 h respectively: ( E ) Representative western blots of cardiomyocyte hypertrophic markers, MYH7 and ANP; ( F ) and ( G ) shows the pooled quantitative densitometry analysis of MYH7 and ANP respectively, The mean with SEM is presented. biological replicate, n = 3–4 for each condition; * p < 0.05 and ** p < 0.01 all by student’s t-test; ( H ) Immunofluorescence data showing the effect of ET—1 and S6K1i in siFTO-CMs and scr-CMs (alpha-actinin in green and DAPI in blue); ( I ) Measurement of average cell area in µm. 2 quantified using Image J; The mean with SEM is presented. n represents single cell (Scr Un, n = 167; Scr ET-1, n = 116; Scr ET1 + S6K1i, n = 92; siFTO Un, n = 103; siFTO ET-1, n = 94; siFTO ET-1 + S6K1i, n = 166) statistical analysis is performed using one-way ANOVA column analysis * p < 0.05; *** p < 0.001; **** p < 0.0001

Article Snippet: The Taqman Universal PCR master mix (Cat. No. 4304437) and the specific qPCR primers for AKT1S1 (TaqMan Gene expression Assay ID Hs00982883_m1; AKT1S1); TFEB ( TaqManTM Gene Expression Assay Hs00292981_m1; TFEB); and FTO (Taqman Gene expression Assay ID Hs01057143_m1; FTO) were purchased from Invitrogen.

Techniques: Transfection, Concentration Assay, TUNEL Assay, Staining, Western Blot, Activity Assay, Marker, Incubation, Immunofluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Deregulation of m6A-RNA methylation impairs adaptive hypertrophic response and drives maladaptation via mTORC1-S6K1-hyperactivation and autophagy impairment

doi: 10.1186/s12964-025-02509-0

Figure Lengend Snippet: Proposed model of regulatory effects of FTO dependent m6A methylation in cardiac remodeling: Under acute stress, FTO dependent m6A methylations cordially regulate mTORC1-S6K1 pathway to conduct cardiac hypertrophy which is the early stage of adaptive cardiac remodeling. However, under chronic stress conditions, FTO is diminished and m6A network is imbalanced; mechanistically, FTO deficiency increases mTORC1-S6K1 hyperactivation and defective autophagy subsequently inducing cardiac atrophy and apoptosis, eventually heart failure

Article Snippet: The Taqman Universal PCR master mix (Cat. No. 4304437) and the specific qPCR primers for AKT1S1 (TaqMan Gene expression Assay ID Hs00982883_m1; AKT1S1); TFEB ( TaqManTM Gene Expression Assay Hs00292981_m1; TFEB); and FTO (Taqman Gene expression Assay ID Hs01057143_m1; FTO) were purchased from Invitrogen.

Techniques: Methylation